Enrichment and detection of circulating tumor cells by immunomagnetic beads and flow cytometry
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ORIGINAL RESEARCH PAPER
Enrichment and detection of circulating tumor cells by immunomagnetic beads and flow cytometry Lei Hu . Xueran Chen . Meng Chen . Jinman Fang . Jinfu Nie . Haiming Dai
Received: 25 May 2020 / Accepted: 14 September 2020 Ó Springer Nature B.V. 2020
Abstract Objective The purpose of the article is to establish a quick enrichment and detection method using immunomagnetic beads and flow cytometry to analyze circulating tumor cells (CTCs) in the peripheral blood. Results After incubation with CD326-PE and CD45APC antibodies, more than 60% MCF7 cells in M-Buffer could be detected while less than 10% of the same cells could be detected by flow cytometry (FCM) if spiked into blood. However, in combination Lei Hu and Xueran Chen have contributed equally to this work.
with CD326 and CD45 immunomagnetic beads, detection rate of MCF7 cells in blood reached 57%. For circulating tumor cells, enrichment by CD326 and CD45 immunomagnetic beads improve the detection rate from nearly undetectable to more than 24.14%. Conclusions Live CTCs in peripheral blood can be effectively and sensitively detected by using a combination of immunomagnetic beads (CD45 and CD326) and flow cytometry. Keywords Circulating tumor cells Flow cytometry Fluorescence antibodies Immunomagnetic beads
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-03007-8) contains supplementary material, which is available to authorized users. L. Hu X. Chen M. Chen J. Fang J. Nie (&) H. Dai (&) Anhui Province Key Laboratory of Medical Physics and Technology, Institute of Health & Medical Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China e-mail: [email protected]
M. Chen University of Science and Technology of China, Hefei 230026, China J. Nie Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China
H. Dai e-mail: [email protected] L. Hu X. Chen M. Chen J. Fang J. Nie H. Dai Hefei Cancer Hospital, Chinese Academy of Sciences, 350 Shushanhu Road, Hefei 230031, Anhui, China
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Biotechnol Lett
Introduction High patient mortality of tumor is mainly due to metastasis (Talmadge and Fidler 2010). Circulating tumor cells (CTCs), which are detached cells slough off the edges of a solid tumor and swept away by the bloodstream or lymphatic system, represent the primary cause of metastasis formation (Alix-Panabieres et al. 2012; Mego et al. 2011). CTCs can remain loose in circulation, cluster together as they travel or settle themselves in new tissues (Alix-Panabieres and Pantel 2014). Because of the same origin between CTCs and primary tumor, CTCs often hold the genetic information of the primary tumor. Thus, CTCs are also used to evaluate the genetic information of tumors. As a promising liquid biopsy, the acquisition of specimens for CTC detection is more convenient, less traumatic, less painful, and more repeatable than the traditio
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